Development of salt-tolerant lines of Malaysian indica rice (Oryza sativa L. cv. MR219) using tissue culture approach

Rice is one of the most important staple foods for human. However, millions hectares of rice land in the South and Southeast Asia were left uncultivated and/or grown with very low yields due to salinity. Due to this, development and production of salt-tolerant rice seed as apart of integrated management practices is needed. By using tissue culture approach, salt-tolerant lines of Malaysia indica rice cv. MR219 were produced. This work started with production and propagation of MR219 callus. Then, callus was sub-cultured separately on MS media supplemented with 2 mg/L 2,4-D and different concentration of NaCl (0, 50, 100, 200, and 300 mM NaCl) to produce salt-tolerant MR219 callus. Screening and selection of salt-tolerant MR219 callus were conducted using morphological and biochemical markers which are total proline content, total soluble sugar, lipid peroxidase and the activity of ascorbate peroxidase and catalase. At regeneration of salt-tolerant plantlets, selected salt-tolerant callus was sub-cultured on MS media supplemented with 2 mg/L kinetin and 1 mg/L BAP for shoot induction, followed by sub-cultured in MS media supplemented with 0.5 mg/L BAP, 1 mg/L kinetin, 1 mg/L IBA and 0.5 mg/L NAA for root formation. At acclimatization stage, MR219 plantlets from 50 mM NaCl found survived and transferred to pots containing paddy soil. These plantlets is called First generation (F1) salt-tolerant MR219. After 70 days, seeds of F1-salt-tolerant MR219 lines was successfully obtained. The grain characteristics of mother plant and F1-salt-tolerant MR219 lines were compared. Germination capability of F1-salt-tolerant MR219 seed in saline showed that seeds of F1-salt-tolerant MR219 able to germinate and growth in 50 -100 mM NaCl. As conclusion, salt-tolerant MR219 rice was produced in vitro and have potential to be commercialized. The protocol to produce salt-tolerant rice can be used to produce other salt-tolerant plant

Physico-chemical parameters and trophic status of an acidic lake in Hutan Simpan Ayer Hitam, Puchong

Relationship between physico-chemical parameters including pH, Temperature, total dissolved solid, dissolved oxygen, light intensity, ammonia, nitrate, silica, and phosphate with microalgae diversity were studied. Water sampling water was conducted biweekly for 4 months from October 2015 to February 2016 for 15 weeks in North Lake of Hutan Simpan Ayer Hitam Puchong. Physico-chemical parameters were determined at site and laboratory. Data analysis including Pearson Correlation Analysis, Principal Component Analysis, and Canonical Correspondence Analysis were carried out. Green algae, Staurastrum sp. and Chlamydomonas have a positive correlation with pH and total dissolved solid. Cosmarium spp. have a positive correlation with dissolved oxygen and have association with increased pH and ammonia. Diatom, Navicula sp. have high negative correlation with pH and have association with increased of silica. Dinoflagellates, Gymnodinium sp. and Gonyaulax apiculata both have high negative correlation with pH. Gymnodinium sp. have association with increased phosphate and temperature. Gonyaulax apiculata have a positive correlation with temperature. Based on research, this lake were slightly acidic but remains oligotrophic North Lake of Hutan Simpan Ayer Hitam Puchong are acidic but oligotrophic due to pH in range of acidic condition but below the boundary of phosphate concentration from 0.01mg/L to 0.075mg/L suggested by USEPA (2002) in aquatic systems

Current status of algae biodiversity and succession in North Lake of Hutan Simpan Ayer Hitam, Puchong

This study presents data on the diversity and succession of algae in North Lake, Hutan Simpan Air Hitam (HSAH) as well as its relationship with chemical and physical factors. Algae samples were collected weekly from three different sites in North Lake started from October 2015 until January 2016 (15 weeks). Physico-chemical parameters such as pH, temperature, light, dissolved oxygen (DO), total dissolved solid (TDS) and nutrients statuses were measured simultaneously. A total of 18 species were recorded comprising Bacillariophyta (3 species), Charophyta (4 species), Chlorophyta (4 species), Cyanobacteria (2 species), Dinophyta (4 species), and Ochrophyta (1 species). The highest total density of algal cells were phyla Dinophyta (24%) followed by Charophyta (22%), Bacillariophyta (19%), Ochrophyta (18%), Cyanophyta (10%) and Chlorophyta (7%). Dinobryon sertularia was the most frequent species found throughout the 15 weeks of sampling and became dominant in week 3, 4, 6, 10 and 13; whereas Staurastrum sp. achieved the highest algal counts in week 1 and 2. Gonyaulax apiculata was dominant during week 5, 8, 14 and 15. The Shannon-Weaver diversity Index, H’ classified North Lake as being moderately polluted, whereas the pH values indicates that the lake was quite acidic by having pH lower than 7 and ranged between 4.28 and 5.57. The occurrence of some phytoplankton groups may be influenced by pH, as has been found in acidic natural lakes and also in extremely acidic lakes around the world. In addition, other physico-chemical factors may also give impact on algae structure and community

Biodiversity and succession of freshwater algae in Hutan Simpan Ayer Hitam Forest Reserve, Puchong Selangor

The succession and dominant of freshwater algae were studied in the north lake of Ayer Hitam Forest Reserve. Sampling was performed for thirteen weeks, from the end of the November 2013 to the end of the February 2014. Water sampling for biological analysis was done during the thirteen weeks. The structural community was determined by population density, class of algae and diversity indices. The community of freshwater algae consisted of 15 species from 5 different phyla which are Ochrophyta, Dinophyta, Charophyta, Cyanophyta and Chlorophyta. The Dinophyta had the highest total density in the thirteen weeks and two species from three species of Dinophyta were the most conspicuous species in the succession which are Gonyaulax apiculata and Gymnodinium palustre. The dominance of Dinophyta was during week 2, 3, 6, 7, 10 and 11. Dinobryon sertularia from phylum Ochrophyta had the highest density and being dominant during week 1, 4 and 5. For the Cyanophyta, Anabaena subcylindrica had the highest density as compared to the other species of Cyanophyta. Anabaena subcylindrica was being dominant during week 8, 9 and 13. For diatom Navicula sp. (Ochrophyta), the highest density was obtained during week 12. The Shannon-Weaver index had the highest during week 12 and the lowest during week 4. The highest Evenness index occurred during week 3 and the lowest during week 4. The pattern of succession and dominant of freshwater algae in a north lake of AHFR was associated with the physico-chemical parameter especially nutrient status, pH, temperature and light intensity

Bioeconomic study a marine phytoplankton Chaetoceros calcitran by using commercial and formulated fertilizer

Studies were carried out on two different aspect of marine diatom, Chaetoceros calcitran. First is the measure of cell growth base on number of cells and dry weight in different culture media such as LCP, LCP + clewat 32, Japanese Media and Conway Media as reference. The second study is looking at the effect of growth in different tank such as glass tank, conical fiberglass tang and rectangular fiberglass tank. All the culture of Chaetoceros calcitran, was carried out in outdoor tank, salinity of 23.25 % and ambient air temperature of 27-36 °C. Throughout the study, the weather is sunny. Results shows that Chaetoceros calcitran cultured in Conway media has the highest cell at 1.843 x 106 cells (maximum cell number) and 0.870 g/L (maximum dry weight). The Cultivation cost for Conway media and LCP were RM 41.227 and RM 74.833 respectively. The results from second showed growth are in the glass tank which is 1.687 x 106 cell/ml (maximum cell growth) and 0.783 g/L (maximum dry weight). Cultivation in a glass tank also showed that the lowest production cost at RM 45.191 /g dry weight compared to the fiber conical tank and rectangular fiber glass tank

Algal communities analysis as an assessment of the trophic status of Ayer Hitam Forest Reserve

Studies of algal biodiversity in relation to nutrient status of water bodies in Malaysia are lacking. In Ayer Hitam Forest Reserve (AHFR), succession of the freshwater algae and physico-chemical parameters were studied in 16 weeks starting from November 2014 until February 2015. Identification and enumeration of algae cells were done using Neubauer chamber (hemocytometer), whereas physico-chemical parameters including pH, water temperature, light intenstiy, dissolved oxygen, total dissolved solid were determined in situ. Parameters such as ammonia, nitrate, phosphate and silica were measured in the laboratory. Pearson Correlation Analysis principal component analysis (PCA) and canonical component analysis (CCA) were used to establish correlation with the physico-chemical parameters such as light intensity, temperature, pH, total dissolved solid, dissolved oxygen and nutrients status including the concentration of ammonia, nitrate, phosphate and silica. Dominating algae species found were Dinobryon sertularia, Gymnodinium palustre, Gonyaulax apiculata and Navicula sp. . Gonyaulax apiculata and Dinobryon sertularia were found to have correlation with total dissolved solid, nitrate, phosphate and light intensity. Shannon-weaver and evenness indices were used to measure the species diversity at the lake and indicated that the lake was low in nutrients (oligotrophic lake). The species richness and diversity was quite stable and the water in the lake was indicated as moderately polluted. Due to the acidic condition of the lake, only selected algae species tolerated, survived and dominated the lake

A metagenome for lacustrine Cladophora (Cladophorales) reveals remarkable diversity of eukaryotic epibionts and genes relevant to materials cycling

Periphyton dominated by the cellulose-rich filamentous green alga Cladophora forms conspicuous growths along rocky marine and freshwater shorelines worldwide, providing habitat for diverse epibionts. Bacterial epibionts have been inferred to display diverse functions of biogeochemical significance: N-fixation and other redox reactions, phosphorus accumulation, and organic degradation. Here, we report taxonomic diversity of eukaryotic and prokaryotic epibionts and diversity of genes associated with materials cycling in a Cladophora metagenome sampled from Lake Mendota, Dane Co., WI, USA, during the growing season of 2012. A total of 1,060 distinct 16S, 173 18S, and 351 28S rRNA operational taxonomic units, from which >220 genera or species of bacteria (~60), protists (~80), fungi (6), and microscopic metazoa (~80), were distinguished with the use of reference databases. We inferred the presence of several algal taxa generally associated with marine systems and detected Jaoa, a freshwater periphytic ulvophyte previously thought endemic to China. We identified six distinct nifH gene sequences marking nitrogen fixation, >25 bacterial and eukaryotic cellulases relevant to sedimentary C-cycling and technological applications, and genes encoding enzymes in aerobic and anaerobic pathways for vitamin B12 biosynthesis. These results emphasize the importance of Cladophora in providing habitat for microscopic metazoa, fungi, protists, and bacteria that are often inconspicuous, yet play important roles in ecosystem biogeochemistry

Bioeconomic study of Chlorella vulgaris in outdoor tank culture by using different types of fertilizer

Four different culture mediums that were Bold Basal, commercial fertilizer, modified fertilizer and modified Bold Basal were used to cultivate Chlorella sp. in outdoor tanks. The modified fertilizer was formulated according to the NPK ratio in Bold Basal medium (control). Clewat 32 (0.01 g/L) was used in modified Bold Basal to replace the micronutrient in Bold Basal medium. The growth of C. vulgaris in outdoor culture was determined daily by cell count and dry weight every two days. The result of this study shows that, culture in modified fertilizer has no effect in improving the total production in outdoor culture. Meanwhile, culture in Bold Basal medium has relatively higher productivity (1.14 g/m2/day) at its optimum if compared to modified Bold Basal medium at 1.08 g/m2/day. Thus, the production cost per gram in 100 L outdoor tank culture was less expensive for Bold Basal medium which is about RM 21.13/g but moderately higher in modified fertilizer (RM 26.75/g). Bioeconomic studies revealed the weakness particularly with respect to the biological component. Possible weaknesses in the biological component are low productivity, unsuitable strain of Chlorella sp., weather, culture technique and scale of production

New records for freshwater dinoflagellates: Peridinium sp. and Gonyaulax sp. discovered in North Lake of Hutan Simpan Ayer Hitam, Puchong

In Malaysia, there are very less emphasis given on freshwater dinoflagellates, hence less information are available for freshwater species which also plays a part in ecological functions. This study focused on the diversity and abundance of dinoflagellate species found at the oligotrophic lake of North Lake of Hutan Simpan Ayer Hitam, Puchong. Dinoflagellates are flagellated protist that could be photosynthetic and hetorotropic in both freshwater and marine waters. They are the crucial primary producer being second only to the diatoms among the micro-phytoplankton in the 10μm to 200μm. This research focused on the morphological and taxonomic studies of those dinoflagellates discovered through microscopical identification using scanning electron microscope up to 3500 X magnification. This study revealed out the existence of different species dinoflagellates believed to belong to two different genus of Peridinium (order Peridiniales) and Gonyaulax (order Gonyaulacales). Three species of Peridinium and 2 species of Gonyaulax were identified. Morphological and taxonomic identification were done using their thecal plate counts and different suture width and pattern. Peridinium has a distinctive characteristics of having 2 antapical plates (2’’’’) while Gonyaulax to possess single antapical plate (1’’’’). The actual plate formula for species belonging to order Peridiniales is 4’, 2-3a, 7’’, 5’’’, 2’’’’ while order Gonyaulacales is 4’, 5’’-6’’,6C, 5S (sulcal plates), 6’’’,1’’’’. The width of the suture for Peridinium sp. varies from approximately ~1μm to 2μm while for Gonyaulax sp, it accounted for ~1 μm to 5.5 μm. As a conclusion, the study on Dinoflagellates belonging to the phylum of Dinophyceae from the North Lake of Hutan Simpan Ayer Hitam brought a new information on the diversity of dinoflagellates thriving in this lake which could be utilized as a reference if there were freshwater algal blooms occurring in Malaysia in order to trace the organism causing it

The cell structure and growth rate of three species of marine microalgae (Chlorella vulgaris, Tetraselmis chuii and Isochrysis sp.) before and after cryopreservation

Three species of marine phytoplankton Chlorella vulgaris, Tetraselmis chuii and Isochrysis sp. were cultured using Conway media in 20 °C with the light intensity of 90 μmol m-2s-1 and a photoperiod of 12:12 h L/D cycle. Cells were harvested in stages at logarithmic and stationary phases. Equilibration time for cyroprotectant exposure was 10 to 20 minutes before the samples was placed in the programmable freezer for a controlled rate of cooling at -1 °C /min. The temperature was reduced uniformly from 200C until it reaches -40 °C where the samples were maintained at this temperature for 30 minutes before direct immersion into liquid nitrogen at -196 °C. Samples were stored in liquid nitrogen for one week. Percentage of viable cells was relatively higher in the stationary phase for the three species that is 95.13 % for Chlorella vulgaris, 79.09 % for Isochrysis sp. and 69.36 % for Tetraselmis chuii. The growth rate of viable cells for Chlorella vulgaris a both logarithmic and stationary phases and Tetraselmis chuii at logarithmic phase showed insignificant differences (p > 0.05) before and after cryopreservation. Tetraselmis chuii of stationary phase and Isochrysis sp. of both phases gave significant differences (p < 0.05) for growth rate before and after cryopreservation due to photo-oxidation. The cell structure after cryopreservation showed disorganization and abnormality in the ultra-structure but this did not seem to affect the growth rate of the viable cells